CN111925962A - Pediococcus ethanolicus capable of producing mannitol and diallyl disulfide - Google Patents
Pediococcus ethanolicus capable of producing mannitol and diallyl disulfide Download PDFInfo
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Abstract
The invention discloses a pediococcus ethanolicus capable of producing mannitol and diallyl disulfide, and belongs to the technical field of microorganisms. The invention screens out a strain of Pediococcus aeolicus which has high growth speed, can inhibit mould and microzyme, can produce organic acid, mannitol and diallyl disulfide with high yield, and is specifically represented by the following steps: inoculating the ethanol-resistant Pediococcus fasciatus into a pickle raw material for fermentation for 12 hours, so that the viable count of lactic acid bacteria in the pickle can reach 8.5lgCFU/mL, the total viable count of mould and yeast in the pickle can be reduced to 1.6lgCFU/mL, the pH value of the pickle can be reduced to 3.5 by fermentation for 48 hours, and the total acid content in the pickle can reach 0.6g/100g, the mannitol content can reach 565.18mg/100g and the diallyl disulfide content can reach 793.10 mu g/kg by fermentation for 96 hours.
Description
Technical Field
The invention relates to a pediococcus ethanolicus strain capable of producing mannitol and diallyl disulfide, and belongs to the technical field of microorganisms.
Background
The pickle is a fermented food prepared by taking fresh vegetables as raw materials, adding seasonings, and carrying out lactic acid fermentation in an anaerobic environment by utilizing microorganisms attached to the vegetables or additionally added lactic acid bacteria leaven. In general, vegetables rich in fiber can be used as pickles, such as cabbage, chinese cabbage, carrot, white radish, garlic, green onion, cucumber, onion, and Korean vegetable.
The production of pickles mainly relies on the fermentation of lactic acid bacteria. Research shows that during the fermentation process of pickled vegetables, dominant strains of pickled vegetables such as leuconostoc mesenteroides, streptococcus mutans, pediococcus ethanolici, lactobacillus plantarum and pediococcus pentosaceus can generate substances such as organic acid and the like, inhibit the growth of other mixed bacteria such as mold and yeast, and simultaneously generate flavor substances such as mannitol and diallyl disulfide which are decisive for the course of the special flavor of the pickled vegetables (see references' Wisselink H, Weusthuis R, Eggink G, et al. manitol production by lactic acid bacteria: A review [ J ]. International Dairy Journal, 2002, 12(2): 151-.
Therefore, obtaining lactic acid bacteria capable of highly producing flavor substances such as organic acid and mannitol, diallyl disulfide and the like is very key to ensuring the success rate of pickle production and improving the flavor of pickle.
However, there are not many dominant strains of kimchi fermentation such as Leuconostoc mesenteroides, Streptococcus mutans, Pediococcus ethanolicus, Lactobacillus plantarum, Pediococcus pentosaceus and the like that can produce organic acids and flavor substances such as mannitol, diallyl disulfide and the like at high yield, and for example, Loesche et al found that Streptococcus mutans can produce only 0.1 to 0.3mg of mannitol in a reaction containing 1.5% (w/v) sucrose (see, specifically, reference "Loesche WJ, Kornman KS. production of mannose by streptococcus mutans [ J ]. Archives of Oral Biology, 1976, 21: 551-553").
Therefore, it is urgently needed to find more kimchi fermentation dominant strains capable of highly producing organic acids and flavor substances such as mannitol, diallyl disulfide and the like so as to ensure the success rate of kimchi production and improve the kimchi flavor.
Disclosure of Invention
[ problem ] to
The technical problem to be solved by the invention is to provide a Pediococcus ethanoliduranus strain capable of highly producing organic acid, mannitol and diallyl disulfide.
[ solution ]
In order to solve the technical problem, the invention provides a Pediococcus ethanolidus (VCQWZ 3-L7), wherein the VCQWZ3-L7 is preserved in Guangdong provincial microorganism strain preservation center in 30 days 07 and 2020, and the preservation number is GDMCC No.61112, and the preservation address is No. 59, 5 th of Dazhou institute No. 100 Mr. of Pieli Zhonglu, Guangzhou.
The Pediococcus ethanolidianus VCQWZ3-L7 is obtained by separating a pickled vegetable water sample from Chongqing city, the 16S rDNA sequence of the strain is shown as SEQ ID NO.1 through sequencing analysis, and the sequence obtained through sequencing is compared with the nucleic acid sequence in NCBI, so that the strain is shown to be the Pediococcus ethanolidianus and is named as the Pediococcus ethanolidianus VCQWZ 3-L7.
The invention also provides a method for producing mannitol and/or diallyl disulfide, which comprises the steps of inoculating the Pediococcus ethanolicus VCQWZ3-L7 into a fermentation medium for fermentation to obtain a fermentation broth, and extracting mannitol and/or diallyl disulfide from the fermentation broth.
The invention also provides the use of the Pediococcus ethanolicus VCQWZ3-L7 or the process described above in the production of mannitol and/or diallyl disulfide.
The invention also provides a leaven, which contains the ethanol-tolerant pediococcus VCQWZ 3-L7.
In one embodiment of the invention, the preparation method of the leaven comprises the steps of inoculating the ethanol-resistant pediococcus VCQWZ3-L7 into a culture medium for culture to obtain a culture solution; centrifuging the culture solution to obtain thalli; resuspending the thalli by using a buffer solution and a freezing protective agent to obtain a resuspension solution; and (4) carrying out vacuum freeze drying on the heavy suspension to obtain the leavening agent.
In one embodiment of the present invention, the leavening agent is a kimchi leavening agent.
The invention also provides a method for producing pickled vegetables, which comprises the step of inoculating the ethanol-resistant pediococcus VCQWZ3-L7 or the leaven into vegetables for anaerobic fermentation to obtain the pickled vegetables.
In one embodiment of the present invention, the fermentation temperature is 20-30 ℃ and the fermentation time is 3-4 days.
In one embodiment of the invention, the vegetables are white radish, chili pepper, garlic and ginger.
The invention also provides pickled vegetables which are produced by using the method.
The invention also provides the application of the Pediococcus Ethocolchicus VCQWZ3-L7 or the leavening agent or the method in the production of pickled vegetables.
The invention also provides a fungal inhibitor, which contains the Pediococcus ethanolicus VCQWZ 3-L7.
In one embodiment of the invention, the fungus is a mold and/or a yeast.
The invention also provides a method for inhibiting fungi, which is to culture the pediococcus ethanolicus VCQWZ3-L7 or the fungal inhibitor by adding the pediococcus ethanolicus VCQWZ3-L7 or the fungal inhibitor into a culture system containing fungi or fungal spores.
In one embodiment of the invention, the fungus is a mold and/or a yeast.
The invention also provides the application of the Pediococcus ethanolicus VCQWZ3-L7 or the leavening agent or the method in inhibiting fungi, which is not aimed at diagnosing and treating diseases.
In one embodiment of the invention, the fungus is a mold and/or a yeast.
[ advantageous effects ]
The invention screens out a strain of Pediococcus ethanolidus (VCQWZ 3-L7), the growth speed of the Pediococcus ethanolidus VCQWZ3-L7 is fast, the fungus and the yeast can be inhibited, and the organic acid, the mannitol and the diallyl disulfide can be produced with high yield, which is specifically embodied as follows:
(1) inoculating the ethanol-resistant Pediococcus fasciatus VCQWZ3-L7 into the pickle raw material for fermentation for 12 hours, so that the viable count of the lactic acid bacteria in the pickle can reach 8.5 lgCFU/mL;
(2) inoculating the Pediococcus Etorhybridus VCQWZ3-L7 into the pickle raw material for fermentation for 24h, so that the total viable count of mould and yeast in the pickle can be reduced from 5.5lgCFU/mL to 1.6 lgCFU/mL;
(3) inoculating the ethanol-resistant Pediococcus fasciatus VCQWZ3-L7 to the pickle raw material for fermentation for 48 hours, so that the pH value of the pickle can be reduced to 3.5, and the total acid content in the pickle can be up to 0.6g/100g after fermentation for 96 hours;
(4) inoculating the ethanol-resistant Pediococcus fasciatus VCQWZ3-L7 into a pickle raw material for fermentation for 96 hours, so that the content of mannitol in the pickle can reach 565.18mg/100 g;
(5) the ethanol-tolerant Pediococcus fasciatus VCQWZ3-L7 is inoculated into a pickle raw material for fermentation for 96 hours, so that the diallyl disulfide content in the pickle can reach 793.10 mug/kg,
therefore, the Pediococcus ethanolicus VCQWZ3-L7 has extremely high application prospects in the aspects of producing mannitol and/or diallyl disulfide, producing pickles, inhibiting fungi and the like.
Biological material preservation
A strain of Pediococcus ethanolidus (Pediococcus ethanolidus) VCQWZ3-L7, which is classified and named as Pediococcus ethanolidus, is preserved in Guangdong provincial microorganism strain preservation center in 30.07-2020, and has the preservation number of GDMCC No.61112 and the preservation address of No. 59, 5 th of Dazhou No. 100 Md.
Drawings
FIG. 1: colonial characteristics of Pediococcus Ethocolchicus VCQWZ 3-L7.
FIG. 2: the thallus characteristics of Pediococcus ethanol-resistant VCQWZ 3-L7.
FIG. 3: pH value of pickle obtained by fermenting different ethanol-resistant pediococcus.
FIG. 4: the total acid content of the pickles obtained by fermenting different ethanol-resistant pediococcus.
FIG. 5: viable count of lactic acid bacteria in pickles obtained by fermenting different ethanol-resistant pediococci.
FIG. 6: the total viable count of mould and yeast in the pickle obtained by fermenting different ethanol-resistant pediococcus.
FIG. 7: the mannitol content in the pickle obtained by fermenting different ethanol-resistant pediococcus.
FIG. 8: the content of diallyl disulfide in the pickles obtained by fermenting different ethanol-tolerant pediococcus.
Detailed Description
The invention is further elucidated with reference to a specific embodiment and a drawing.
The white radish, chili pepper, garlic, ginger, salt and spices referred to in the following examples were purchased from the tin-free European supermarket.
The media involved in the following examples are as follows:
MRS solid medium: 10g/L of peptone, 10g/L of beef extract, 20g/L of glucose, 2g/L of sodium acetate, 5g/L of yeast powder and 2g/L, K of diammonium hydrogen citrate2HPO4·3H2O 2.6g/L、MgSO4·7H2O 0.1g/L、MnSO4·H2O0.05 g/L, Tween 801mL/L, agar powder 15g/L, cysteine hydrochloride 0.5 g/L.
MRS liquid medium: 10g/L of peptone, 10g/L of beef extract, 20g/L of glucose, 2g/L of sodium acetate, 5g/L of yeast powder and 2g/L, K of diammonium hydrogen citrate2HPO4·3H2O 2.6g/L、MgSO4·7H2O 0.1g/L、MnSO4·H2O0.05 g/L, Tween 801mL/L and cysteine hydrochloride 0.5 g/L.
The detection methods referred to in the following examples are as follows:
the detection method of the pH value comprises the following steps: weighing 5g of pickle samples, adding 15mL of distilled water into the pickle samples, homogenizing and filtering to obtain juice; 5mL of the juice was collected, and pH of the kimchi was measured by a pH meter.
The detection method of the total acid content comprises the following steps: weighing 5g of pickle samples, adding 15mL of distilled water into the pickle samples, homogenizing and filtering to obtain juice; taking 5mL of juice, and determining the total acid content of the pickle according to the national standard GB/T12456-2008 'determination of total acid in food'.
The detection method of the viable count of the lactic acid bacteria comprises the following steps: weighing 5g of pickle samples, adding 15mL of distilled water into the pickle samples, homogenizing and filtering to obtain juice; taking 5mL of juice, and determining the viable count of the pickle lactobacillus according to the national standard GB 4789.35-2016 lactic acid bacteria inspection for food safety national standard food microbiology.
The detection method of viable count of mould and yeast comprises the following steps: weighing 5g of pickle samples, adding 15mL of distilled water into the pickle samples, homogenizing and filtering to obtain juice; taking 5mL of juice, counting mould and yeast in the pickle by using a Bengal red culture medium, distinguishing according to different morphological characteristics of the mould and the yeast, and specifically referring to the national standard GB 4789.15-2016 (food safety national standard food microbiology test mould and yeast count).
The detection method of the mannitol content comprises the following steps:
weighing 5g of pickle samples, adding 15mL of distilled water into the pickle samples, homogenizing and filtering to obtain juice; centrifuging 1mL of juice at 10000g for 10min, and collecting supernatant for later use; taking 20 mu L of supernatant, adding 15 mu L of ribitol with the concentration of 0.2mg/mL serving as an internal standard substance into the supernatant to obtain a sample; drying the sample in a vacuum freeze concentration instrument for 2 hours, and performing derivatization treatment after the water in the sample is completely volatilized to obtain a sample to be detected; taking 100 mu L of sample to be detected, and adding the sample to be detected into a GC sample injection bottle for GC-MS detection; performing substance qualitative determination by searching the spectrogram measured by GC-MS in NIST2001 standard spectral library and comparing the standard substances, and quantifying by using an area normalization method to obtain the content of mannitol in the pickle sample;
wherein, the derivatization treatment comprises the following steps: adding a sample into a centrifuge tube, adding 80 mu L of methoxylamine hydrochloride solution with the concentration of 20mg/mL into the centrifuge tube, preserving the temperature of the centrifuge tube in a metal bath at 37 ℃ for 90min, adding 40 mu L of N-methyl-N- (trimethylsilyl) -trifluoroacetamide (MSTFA) containing 1% (v/v) trimethylchlorosilane into the centrifuge tube, preserving the temperature of the centrifuge tube in the metal bath at 40 ℃ for 30min, and centrifuging the centrifuge tube at 10000g for 5min to obtain a sample to be detected;
gas chromatography conditions: rtx-5MS caliper column (30m × 0.25mm × 0.25 μm) was used for separation, the sample injection volume was 1 μ L, the split ratio was 10:1, the carrier gas was helium, the flow rate was 1mL/min, the sample injection port temperature was 240 ℃, the column initial temperature was 70 ℃, the temperature was raised to 230 ℃ at 5 ℃/min, the temperature was raised to 320 ℃ at 90 ℃/min, and the temperature was maintained for 5 min.
Mass spectrum conditions: the temperatures of the transmission line and the ion source are respectively 280 ℃ and 320 ℃, the temperature of the quadrupole is 150 ℃, and the mass spectrum scanning range m/z is 33-600.
The detection method of the content of the diallyl disulfide comprises the following steps:
taking 5g of pickle sample, adding the pickle sample into a 20mL screw mouth sample injection bottle, and adding 2 mu L of methyl heptanoate methanol solution with the concentration of 0.05mg/mL into the screw mouth sample injection bottle as an internal standard to obtain a sample; adding a sample into a sample feeding bottle, incubating the sample feeding bottle at 50 ℃ for 5min, inserting an extraction head (DVB/CAR/PDMS) into the sample feeding bottle, extracting at 50 ℃ for 30min, desorbing at 240 ℃ for 2min, and performing GC-MS analysis; performing substance qualitative determination by searching the spectrogram measured by GC-MS in NIST2001 standard spectral library and comparing the standard substances, and quantifying by using an area normalization method to obtain the content of diallyl disulfide in the pickle sample;
wherein, the gas chromatography conditions are as follows: separating with Rtx-WAX capillary (30m × 0.25mm × 0.25 μm), with sample volume of 1 μ L, split ratio of 10:1, flow rate of 1mL/min, helium as carrier gas, sample inlet temperature of 240 deg.C, column initial temperature of 40 deg.C and holding for 3min, heating to 130 deg.C at 5 deg.C/min, holding for 5min, heating to 155 deg.C at 25 deg.C, and heating to 220 deg.C at 5 deg.C/min and holding for 5 min.
Mass spectrum conditions: the temperatures of the transmission line and the ion source are 220 ℃ and 230 ℃, respectively, the ionization mode EI is realized, the temperature of the four-level bar is 150 ℃, and the mass spectrum scanning range m/z is 30-500.
Example 1: screening, identification and preservation of Pediococcus Ethocolchicus VCQWZ3-L7
The method comprises the following specific steps:
1. screening
Taking pickle water from Chongqing city as a sample, and carrying out gradient dilution by 10 times to 10 times by using sterile physiological saline water-6Then 100 mul of the solution is respectively diluted by 10 times-4、10-5、10-6The diluted solution is coated on a MRS solid culture medium by a flat plate, is inversely cultured for 48 hours at 37 ℃, and the colony morphology is observed and recorded; selecting colonies with different forms on an MRS solid culture medium for streaking separation, and after static culture at 37 ℃ for 48 hours, selecting single colonies with different forms on the MRS solid culture medium again for streaking separation until pure single colonies with consistent forms are obtained; observing colony morphology of a single colony, performing microscopic examination, gram staining and physiological and biochemical characteristic inspection on the obtained colony, selecting strains with small, round, milky, convex and smooth colony morphology, spherical thallus morphology, positive gram staining result and positive catalase test, wherein two strains which are possibly resistant to Pediococcus ethanolicus are obtained and are respectively named as VCQWZ3-L7 (the colony characteristics of VCQWZ3-L7 are shown in figure 1, the thallus characteristics of VCQWZ3-L7 are shown in figure 2) and SCYAPCZN 19;
the microscopic examination method comprises the following steps: taking a clean glass slide, slightly heating the clean glass slide on flame, and cooling the clean glass slide; after cooling, a small drop of sterile water is dripped at the central part of the clean glass slide, and a small amount of thalli on a single bacterial colony is picked by an inoculating loop beside the flame and mixed with the water; burning off redundant thalli on the inoculating loop, and coating the thalli into a uniform thin layer with the diameter of 1cm by using the inoculating loop; after the uniform thin layer on the clean glass slide is naturally dried, heating for 3-4 times on a slow fire for fixing; dripping 1-2 drops of crystal violet solution at the smear position, and dyeing for 1 min; washing off the dye solution with water, and sucking off water on the glass slide with water sucking paper; dripping 1-2 drops of iodine solution on the smear, dyeing for 1min, slightly washing off the dye solution with water, and slightly absorbing water on the glass slide with absorbent paper; dripping 1-2 drops of 95% (v/v) ethanol solution on the smear, decoloring for 5-15 s, slightly washing with water, and slightly absorbing water on the glass slide by using absorbent paper; dripping 1-2 drops of safranin solution at the smear position, dyeing for 2min, drying, and performing microscopic examination by using a microscope to observe the thallus form;
the gram staining method was described in textbook "Industrial microbiology Breeding" (authors: Zhugejian);
the examination method of physiological and biochemical characteristics is described in textbook "handbook of identification of common bacterial systems" (Author: Dongxiu bead).
2. Identification
The genome of the strains VCQWZ3-L7 and SCYAPCZN19 is extracted, 16S rDNA of the strains VCQWZ3-L7 and SCYAPCZN19 is amplified and sequenced (the sequencing is carried out by Huada gene, wherein the nucleotide sequence of the 16S rDNA of the strains VCQWZ3-L7 is shown as SEQ ID NO.1, the primers used for amplification are 27F with the nucleotide sequence shown as SEQ ID NO.2 and 1492R with the nucleotide sequence shown as SEQ ID NO. 3), the sequenced sequences are subjected to nucleic acid sequence alignment in NCBI, and the strains VCQWZ3-L7 and SCYAPCZN19 are both ethanol-resistant sheet cocci which are respectively named as ethanol-resistant sheet cocci (Pediococcus ethanolidans) VCQWZ3-L7 and SCPCQWZN (Pediococcus ethanolidis) VCQWZ 11345-VCQWZ 8236 with the sequence number of the ethanol-resistant sheet cocci (Pediococcus ethanolidis recorded in the organization of SCYAPCZN 11325-8236).
3. Preservation of
Selecting single colonies of Pediococcus Ethicus VCQWZ3-L7 and Pediococcus Ethicus SCYAPCZN19, inoculating in an MRS liquid culture medium, and carrying out static culture at 37 ℃ for 20-24 hours to obtain a bacterial liquid; taking 1mL of bacterial liquid in a bacteria-preserving tube, centrifuging at 6000rpm for 3min, removing supernatant, and suspending bacterial mud in 30% (v/v) glycerol solution and preserving at-80 ℃.
Example 2: production of kimchi
The method comprises the following specific steps:
respectively streaking the Pediococcus ethanolicus VCQWZ3-L7 and the Pediococcus ethanolicus SCYAPCZN19 obtained in the example 1 on an MRS solid culture medium, and culturing for 48 hours at 37 ℃ to obtain single colonies; selecting single colonies, respectively inoculating the single colonies in an MRS liquid culture medium, culturing for 18h at 37 ℃ for activation, and continuously activating for two generations to obtain an activation solution; respectively inoculating the activated liquid into an MRS liquid culture medium according to the inoculation amount of 2% (v/v), and culturing for 18h at 37 ℃ to obtain a bacterial liquid; preparing 100g of white radish, 4g of hot pepper, 6g of garlic, 6g of ginger, 6g of salt, 4g of white granulated sugar and 100mL of water; cleaning radix Raphani, Capsici fructus, Bulbus Allii and rhizoma Zingiberis recens, removing impurities on epidermis, scalding with boiling water instantly, cleaning with sterile water for 3 times, and cutting into pieces of 1cm × 1cm to obtain vegetable pieces; mixing the vegetable blocks, salt, white granulated sugar and water to obtain a pickle raw material; putting the pickle raw materials into a pickle jar, inoculating bacterial liquids obtained by culturing Pediococcus ethanolicus VCQWZ3-L7 and Pediococcus ethanolicus SCYAPCZN19 into the pickle raw materials according to the inoculation amount of 2 percent respectively, sealing the pickle jar, and standing and fermenting the pickle jar at 25 ℃ for 96 hours to obtain the pickle.
During the fermentation process, the kimchi is sampled every 24 hours, and the pH value of the kimchi is detected (see detection result in fig. 3), and the content of total acid in the kimchi (see detection result in fig. 4), the number of viable bacteria of lactic acid bacteria (see detection result in fig. 5), and the number of total viable bacteria of mold and yeast (see detection result in fig. 6) are detected.
And sampling the pickled vegetables during fermentation for 96h, and detecting the content of mannitol (shown in a detection result in figure 7) and the content of diallyl disulfide (shown in a detection result in figure 8) in the pickled vegetables.
As can be seen from fig. 3, the pH of the kimchi fermented by pediococcus ethanolici VCQWZ3-L7 decreased faster than that of pediococcus ethanolici SCYAPCZN19, and at 48 hours of fermentation, the pH of the kimchi fermented by pediococcus ethanolici VCQWZ3-L7 decreased to 3.5, while the pH of the kimchi fermented by pediococcus ethanolici SCYAPCZN19 decreased to only 3.85.
As can be seen from FIG. 4, compared with Pediococcus ethanolicus SCYAPCZN19, the content of total acid in the pickled vegetables obtained by fermentation of Pediococcus ethanolicus VCQWZ3-L7 is higher, and when the pickled vegetables are fermented for 96 hours, the content of total acid in the pickled vegetables obtained by fermentation of Pediococcus ethanolicus VCQWZ3-L7 is as high as 0.6g/100g, while the content of total acid in the pickled vegetables obtained by fermentation of Pediococcus ethanolicus SCYAPCZN19 is only 0.5g/100 g.
As can be seen from FIG. 5, the growth rate of Pediococcus ethanolicus VCQWZ3-L7 in kimchi was faster than that of Pediococcus ethanolicus SCYAPCZN19, and the viable count of lactic acid bacteria in kimchi fermented by Pediococcus ethanolicus VCQWZ3-L7 was as high as 8.5lgCFU/mL for 12 hours of fermentation, while the viable count of lactic acid bacteria in kimchi fermented by Pediococcus ethanolicus SCYAPCZN19 was only 7.7 lgCFU/mL.
As can be seen from FIG. 6, compared with Pediococcus ethanolicus SCYAPCZN19, Pediococcus ethanolicus VCQWZ3-L7 has stronger capability of inhibiting the mold and yeast in the pickle, when the pickle is fermented for 24h, the total viable count of the mold and the yeast in the pickle obtained by the fermentation of the Pediococcus ethanolicus VCQWZ3-L7 is rapidly reduced to 1.6lgCFU/mL from 5.5lgCFU/mL before the fermentation, and the total viable count of the mold and the yeast in the pickle obtained by the fermentation of the Pediococcus ethanolicus SCYAPCZN19 is still 3.8 lgCFU/mL.
As can be seen from FIG. 7, compared with Pediococcus ethanolicus SCYAPCZN19, Pediococcus ethanolicus VCQWZ3-L7 has stronger mannitol production capability, and when the fermentation is carried out for 96 hours, the content of mannitol in the pickled vegetables obtained by the fermentation of Pediococcus ethanolicus VCQWZ3-L7 is as high as 565.18mg/100g, while the content of mannitol in the pickled vegetables obtained by the fermentation of Pediococcus ethanolicus SCYAPCZN19 is only 274.10mg/100 g.
As can be seen from FIG. 8, the ability of Pediococcus ethanolicus VCQWZ3-L7 to produce diallyl disulfide was stronger than that of Pediococcus ethanolicus SCYAPCZN19, and the content of diallyl disulfide in the pickles obtained by the fermentation of Pediococcus ethanolicus VCQWZ3-L7 was as high as 793.10. mu.g/kg while the content of diallyl disulfide in the pickles obtained by the fermentation of Pediococcus ethanolicus SCYAPCZN19 was only 308.70. mu.g/kg at 96h of fermentation.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Sequence listing
<110> university of south of the Yangtze river
<120> a strain of Pediococcus ethanolicus capable of producing mannitol and diallyl disulfide
<160> 3
<170> PatentIn version 3.3
<210> 1
<211> 1419
<212> DNA
<213> Pediococcus ethanolicus
<400> 1
tgcagtcgaa cgcactttcg ttaattgatt ttgagatgct tgcatcgaag atgattttaa 60
ctataaagtg agtggcgaac gggtgagtaa cacgtgggta acctgcccag aagtggggga 120
taacacctgg aaacagatgc taataccgca taacaaaatt aaccgcatgg ttgattttta 180
aaagatggct tcggctatca cttctggatg gacccgcggc gtattagcta gttggtgaga 240
taaaggctca ccaaggcagt gatacgtagc cgacctgaga gggtaatcgg ccacattggg 300
actgagacac ggcccagact cctacgggag gcagcagtag ggaatcttcc acaatggacg 360
caagtctgat ggagcaacgc cgcgtgagtg atgaaggctt tcgggtcgta aaactctgtt 420
gttggagaag aacgtgtgtg agagtaactg ctcatgcagt gacggtatcc aaccagaaag 480
ccacggctaa ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg ttatccggat 540
ttattgggcg taaagcgagc gcaggcggtc ttttaagtct aatgtgaaag ccttcggctt 600
aaccgaagaa gtgcattgga aactgggaga cttgagtgca gaagaggaga gtggaactcc 660
atgtgtagcg gtgaaatgcg tagatatatg gaagaacacc agtggcgaag gcggctctct 720
ggtctgcaac tgacgctgag gctcgaaagc atgggtagcg aacaggatta gataccctgg 780
tagtccatgc cgtaaacgat gaatgctaag tgttggaggg tttccgccct tcagtgctgc 840
agctaacgca ttaagcattc cgcctgggga gtacgaccgc aaggttgaaa ctcaaaagaa 900
ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgaagcta cgcgaagaac 960
cttaccaggt cttgacatct tctgctaacc taagagatta ggcgttccct tcggggacgg 1020
aatgacaggt ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg 1080
caacgagcgc aacccttatt attagttgcc agcattaagt tgggcactct agtgagactg 1140
ccggtgacaa accggaggaa ggtggggacg acgtcaaatc atcatgcccc ttatgacctg 1200
ggctacacac gtgctacaat ggacggtaca acgagttgcg agaccgcgag gtttagctaa 1260
tctcttaaaa ccgttctcag ttcggactgc aggctgcaac tcgcctgcac gaagttggaa 1320
tcgctagtaa tcgcggatca gcatgccgcg gtgaatacgt tcccgggcct tgtacacacc 1380
gcccgtcaca ccatgagagt ttgtaacacc caaagccgg 1419
<210> 2
<211> 20
<212> DNA
<213> Artificial sequence
<400> 2
agagtttgat cctggcctca 20
<210> 3
<211> 19
<212> DNA
<213> Artificial sequence
<400> 3
ggttaccttg ttacgactt 19
Claims (10)
1. The Pediococcus ethanolidianus is characterized by being preserved in Guangdong provincial microorganism strain collection center at 30 months 07 and 2020, and the preservation number is GDMCC No. 61112.
2. A method for producing mannitol and/or diallyl disulfide, comprising inoculating the Pediococcus ethanolicus of claim 1 to a fermentation medium for fermentation to obtain a fermentation broth, and extracting mannitol and/or diallyl disulfide from the fermentation broth.
3. Use of Pediococcus ethanolicus according to claim 1 or the method according to claim 2 in the production of mannitol and/or diallyl disulfide.
4. A starter culture comprising Pediococcus ethanol tolerant bacteria of claim 1.
5. A method for producing kimchi, comprising inoculating the Pediococcus ethanol-tolerant Strain of claim 1 or the starter of claim 4 to vegetables and performing anaerobic fermentation to obtain kimchi.
6. A kimchi produced by using the method of claim 5.
7. Use of Pediococcus ethanolicus according to claim 1 or of the starter culture according to claim 4 or of the method according to claim 5 for the production of kimchi.
8. A fungal inhibitor comprising the pediococcus ethanolicus of claim 1.
9. A method for inhibiting fungi, which comprises adding the Pediococcus Ethocolci-resistant according to claim 1 or the fungal inhibitor according to claim 8 to a culture system containing fungi or fungal spores for culture.
10. Use of Pediococcus ethanolicus according to claim 1 or of a starter culture according to claim 8 or of a method according to claim 9 for inhibiting fungi without the purpose of diagnosis or treatment of diseases.
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| CN114752539A (en) * | 2022-05-30 | 2022-07-15 | 四川大学 | Pediococcus ethanol-resistant strain and activation method thereof, microbial inoculum and preparation method and application thereof |
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